空间飞行MG63细胞重力敏感新miRNA HN22的鉴定与功能分析

Identification and Functional Analysis of Gravity-sensitive Novel miRNA HN22 in MG63 during Spaceflight

目的筛选空间飞行引起的MG63成骨细胞差异表达新miRNA并进行功能分析。方法对空间飞行后的MG63成骨细胞small RNA(sRNA)进行测序及生物信息学分析,得到空间飞行导致显著变化的新miRNA。利用地面回转模拟微重力效应实验验证其表达变化情况;采用miRNA转染分析其对成骨细胞分化的影响;采用多种miRNA靶基因预测软件综合预测其靶基因并检测其潜在靶基因的表达情况;通过GO注释和Panther pathway进行靶基因功能分析。结果新miRNA HN22在空间飞行和地面回转模拟微重力条件下均发生显著表达变化;HN22转染显著提高成骨分化相关基因表达。靶基因预测结果分析显示HN22可影响成骨细胞Wnt信号通路与钙黏蛋白信号通路。Wnt信号通路中的Wnt5a与HN22的表达趋势相反,是其潜在的靶基因。结论HN22可响应微重力变化,并靶向Wnt信号通路中的Wnt5a与钙黏蛋白信号通路影响成骨细胞分化。

Objective To screen and identify differential expression of novel miRNAs during spaceflight and analyze their functions.Methods Through the small RNA sequencing and bioinformatics analysis of spaceflight osteoblasts(MG63),differentially expressed novel miRNAs were obtained and their expressions were verified under simulated microgravity.Then the influence of novel miRNA on osteoblasts differentiation were studied by miRNA transfection.Finally,functional analysis of the predicted target genes of novel miRNA was conducted by GO annotation and Panther pathway analysis.Results The novel miRNA HN22 was differentially expressed during space flight and simulated microgravity.HN22 transfection significantly enhanced expression of osteogenic differentiation-related genes and Wnt5 a.Bioinformatic functional analysis showed that HN22 affected Wnt signaling pathway and cadherin signaling pathway in osteoblasts.The expression of HN22 and Wnt5 ashowed negative correlation under simulated microgravity condition.Conclusion HN22 can respond to microgravity and regulate osteoblast differentiation through Wnt signaling pathway(Wnt5 a)and cadherin signaling pathway.